JP3475056B2 - Whole blood cell immunoassay - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a whole blood blood cell immunoassay device.

[0002]

2. Description of the Related Art There is an inflammatory marker as a method of observing the presence or absence of inflammation occurring in the body and its extent and its progress. Typical examples of this inflammatory marker, white blood cell count, erythrocyte sedimentation rate value, acute phase proteins, serum proteins min ethyl, include serum sialic acid was measured by a combination of these, it is help in the diagnosis of inflammation. Among them, the measurement of white blood cell count and C-reactive protein (CRP), which is an acute phase protein, is useful for diagnosing inflammation and infectious diseases. The blood cell counter and the latter were individually measured using an immunoassay device.

[0003]

However, in the case of individually measuring using the above-mentioned blood cell counter and immunoassay device, the sample (specimen) used for the measurement is whole blood in the former case and serum in the latter case mainly. Is. And when obtaining whole blood as a sample, it is necessary to separately collect blood in a state with an anticoagulant and in a state without this, while serum needs to wait for time until blood coagulation and centrifuge. Is not suitable for facilities such as small clinics, clinics, remote clinics, holiday clinics, emergency laboratories, etc. that do not always have specialized laboratory technicians.

On the other hand, there is an immunoassay device that can measure whole blood, but when whole blood is used as a sample, the desired immunoassay item does not exist in blood cells but only in serum or plasma. However, an error occurs due to the variation of the hematocrit value having a relatively large individual difference.

The present invention has been made in view of the above matters, and an object thereof is to provide a whole blood blood cell immunoassay device capable of simultaneously obtaining blood cell count and immune item data.

[0006]

In order to achieve the above object, a whole blood blood cell immunoassay device of the present invention comprises an immunoassay section for performing an immunoassay and a blood cell count and measurement section for performing a blood cell count measurement. The same whole blood sample is used in the measurement unit, and the result of the immunoassay is corrected using the hematocrit value obtained by the blood cell count measurement.

According to the above-mentioned whole blood blood cell immunoassay device, since the blood cell count and the measurement of the immunity item can be carried out at the same time with the whole blood, the sample can be handled only with the whole blood, and it is not necessary to separate the serum, and the blood sample can be collected shortly. The measurement can be started in time, and even a non-specialized inspection engineer can easily perform the measurement. Therefore, it is especially useful for emergency and early diagnosis of inflammation and infectious diseases, and also in small clinics, clinics, remote clinics, holiday clinics, emergency laboratories, etc.
The desired inspection can be performed.

Further, the probe unit section, the arithmetic / control section, the display / output device and the like can be commonly used for the blood cell count measurement and the immunoassay, which is common compared to the conventional individually provided ones. The cost can be reduced as much as possible.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described with reference to the drawings. 1 to 5 show one embodiment of the present invention.

First, FIG. 1 is a perspective view showing an example of a whole blood blood cell immunoassay device of the present invention with a side panel removed, and FIG. 2 shows a general configuration of the whole blood blood cell immunoassay device. FIG. In these figures, 1 is a device case, and a sample setting section 5 for setting a sample container 4 containing whole blood 3 as a sample is formed on the front surface 2 side in a state of being recessed inward. ing. Reference numeral 6 is a measurement key provided in the sample setting section 5.

Below the side surface portion 7 of the device case 1, an immunoassay portion 8 for performing an immunoassay and a blood cell counting and measuring portion 9 for performing a blood cell measurement are viewed from the front side in order from the side closer to the front surface portion 2. The solenoid valve unit 10 is arranged in a straight line and includes a plurality of solenoid valves 10a. Also,
Above the side surface portion 7, a probe unit portion 11 that linearly moves between the sample setting portion 5 and the blood cell counting and measuring portion 9 is provided.

Further, in FIG. 2, reference numeral 12 denotes a fixed-volume dispenser, and 13
Diluent container, 14 hemolysis reagent container 15 is a pump, these 12-15 none is connected to the electromagnetic valve unit 10. Reference numeral 16 is a waste liquid container connected to the pump 15.

The structures of the immunoassay unit 8, the blood cell count measuring unit 9 and the probe unit unit 11 will be described in some detail.

First, the immunoassay unit 8 is configured to measure CRP in this embodiment. That is, in FIG. 2, reference numeral 17 denotes a cell for measuring CRP (hereinafter referred to as a CRP cell), which includes a light irradiation section 17a and a light detection section 17b, and is configured so that the liquid contained therein can be appropriately stirred. Has been done. 18, 1
Reference numerals 9 and 20 denote containers containing reagents used for CRP measurement, and a hemolytic reagent (hereinafter, referred to as R1 reagent), a buffer solution (hereinafter, referred to as R2 reagent), an anti-human CRP sensitized latex immunoreagent (hereinafter, referred to as R3), respectively. (Reagent).

The CRP cell 17 and the reagent containers 18 to 20 are arranged in a straight line with respect to the set position of the sample container 4 in the sample setting section 5.
The solenoid 20 is configured to be opened and closed collectively by a lid 22 that swings up and down by a solenoid 21. Reference numeral 23 is a cooler box provided with an electronic cooler 24 made of, for example, a Peltier element, and in the illustrated example, a reagent R
2 and R3 are housed.

Next, in this embodiment, the blood cell counting and measuring section 9 uses the electric resistance method to determine WBC (white blood cell count), RBC (red blood cell count), PLT (platelet count), MC.
V (red blood cell volume), Hct (hematocrit value), and Hgb (hemoglobin concentration) are measured by the absorptiometric method in the cyanmethemoglobin method. That is, in FIG.
Is a WBC / Hgb hemocytometer cell (hereinafter simply referred to as WBC
Measuring electrode 25 for measuring WBC in a cell)
Light irradiation unit 25 for measuring a, 25b and Hgb
c, and a light receiving section 25d. 26 is RBC / PLT
With a blood cell counter measurement cell (hereinafter simply referred to as RBC cell),
Measuring electrodes 26a, 2 for measuring RBC and PLT
6b is provided. These cells 25 and 26 are CRP
CRP cell 17 and reagent container 18 in measurement unit 8
It is arranged so as to be in line with 20. Also, WB
The C cell 25 also serves as a waste liquid chamber for cleaning a nozzle 33 (described later).

Further, the probe unit section 11 is constructed, for example, as follows. That is, in FIG. 1 and FIG. 2, 27 is a nozzle unit, and the nozzle unit 27 is reciprocated in the horizontal direction by a timing belt 29 provided in the horizontal direction along a vertically extending base member 28. It is configured to be able to. Reference numeral 30 is a motor for driving the timing belt 29, 31 is a pair of guide members for guiding the guided member 32 provided in the nozzle unit 27, and these are attached to the base member 28 via appropriate members. .

A nozzle 33 is attached to a nozzle holder 35 which moves in the nozzle unit 27 in the vertical direction by a timing belt 34. The front end side (lower end side) of the nozzle 33 is configured to be inserted through a nozzle cleaning device 36 provided in the nozzle unit 27 so that the outer periphery of the front end portion is cleaned. A motor 37 drives the timing belt 34. A sensor 38 detects whether the nozzle 33 is at the home position (fixed position).

In FIG. 2, reference numeral 39 denotes a microcomputer as a control / arithmetic device for comprehensively controlling each part of the device and performing various calculations using outputs from the CRP measuring part 8 and the blood cell count measuring part 9. (MCU),
40 is a solenoid valve unit 10 based on a command from the MCU 39,
A driver that sends a drive signal to the motors 30, 37, etc. of the probe unit section 41, a signal processing section 41 that processes output signals from the CRP measurement section 8 and the blood cell count measurement section 9 and sends them to the MCU 39, and 42 is processed by the MCU 39. A device for displaying results obtained by the above, for example, a color display, and 43 is a printer as an output device.

In FIG. 2, the dotted line indicates the flow of the sample 3 and various reagents, the slightly thick one-dot chain line indicates the control signal, and the thin one-dot chain line indicates the signal flow obtained by the measurement. .

The operation of the whole blood blood cell immunoassay device constructed as described above will be described with reference to FIGS. 3 to 5 showing an example of the measurement procedure.

First, the measurement key 6 is turned on (step S
1), the nozzle 33 at the fixed position moves to the position of the R2 reagent (step S2) and sucks the R2 reagent (step S3). After the suction of the reagent, the nozzle 33 moves to the position of the WBC cell 25, the diluted liquid as the cleaning liquid is supplied to the nozzle cleaning device 36, and the outer surface of the nozzle 33 is cleaned.
After that, the nozzle 33 returns to the position of the R2 reagent.

Next, the nozzle 33 moves to the position of the R1 reagent (step S4) and sucks the R1 reagent (step S5). After this reagent suction, the nozzle 33 moves to the position of the WBC cell 25, the diluent is supplied to the nozzle cleaning device 36, and the outer surface of the nozzle 33 is cleaned. After that, the nozzle 33 returns to the position of the R1 reagent.

Then, the nozzle 33 moves to the sample setting position (step S6), and the sample (whole blood) in the sample container 4 is moved.
3 is sucked for CRP measurement (step S7). After this sample suction, the nozzle 33 moves to the position of the WBC cell 25, the diluent is supplied to the nozzle cleaner 35, and the outer surface of the nozzle 33 is cleaned. Then, the nozzle 33 returns to the position of the sample.

Then, the nozzle 33 moves to the position of the CRP cell 17 (step S8), and the sample 3, R1 reagent, R2
The reagent is discharged into the CRP cell 17 (step S9).

Then, the nozzle 33 which has finished the discharge
Moves to the WBC cell 25 position (step S10),
After the sample 3 and the like remaining inside is discharged into the WBC cell 25, the inside and outside thereof are washed with a diluting liquid.

The nozzle 33, which has been cleaned, moves to the sample setting position (step S11), and sucks the sample 3 in the sample container 4 for CBC measurement (step S1).
2). After this sample suction, the nozzle 33 is connected to the WBC cell 25.
After moving to the position, the diluent is supplied to the nozzle cleaning device 36 and the outer surface of the nozzle 33 is cleaned.

The nozzle 33, which has been cleaned, has a WBC
While the sample 3 is discharged into the cell 25, a predetermined amount of the diluent in the diluent container 13 is injected into the WBC cell 25 via the electromagnetic valve unit 10 to perform the primary dilution of the CBC sample (step S13).

The nozzle 33 at the position of the WBC cell 25 is
A predetermined amount of the primary diluted CBC sample is aspirated to generate RB
The CBC 26 is moved to the C cell 26 (step S14), the sucked primary diluted CBC sample is discharged to the cell 26 (step S15), and the diluent in the diluent container 13 is transferred to the RBC via the electromagnetic valve unit 10. A predetermined amount is injected into the cell 26, and the CBC sample is subjected to secondary dilution (step S
16).

After the above primary and secondary dilutions are completed, the hemolytic agent in the hemolytic agent container 14 is transferred to the WBC via the electromagnetic valve section 10.
While a predetermined amount is injected into the cell 25, WBC and Hgb are measured, while RBC and PLT are measured in the RBC cell 26 (step S17), the data at that time is taken into the MCU 39 via the signal processing unit 41. Be done.

When the above measurement is completed, the WBC cell 25 and R
The BC cell 26 is washed with a diluent (step S1).
8).

As described above, steps S10 to S
28, the CBC measurement is performed in the blood cell counting and measuring unit 9, and during this period (for about 60 seconds), the CRP cell 1 is used.
In 7, the hemolytic reaction progresses between the sample 3, the R1 reagent, and the R2 reagent, and the interfering substance is removed.

When the CBC measurement is completed, the nozzle 33 located at the position of the RBC cell 26 moves to the position of the R3 reagent (step S19) and sucks the R3 reagent (step S20). After this reagent suction, the nozzle 33 moves to the position of the CRP cell 17 (step S21), and the R3 reagent is CR.
It is discharged into the P cell 17 (step S22), and the R3 reagent is mixed into the reaction liquid of the sample 3, the R1 reagent, and the R2 reagent.

Then, the liquid is sufficiently stirred in the CRP cell 17 (step S23), an immune reaction occurs and CRP measurement is performed (step S24), and the data at that time is taken into the MCU 39 via the signal processing unit 41. Be done. When the measurement is completed, the CRP cell 17 is washed with a diluent (step S25), and all the measurements are completed (step S26).

In the MCU 39, the RBC (red blood cell count), the red blood cell volume (MC) based on the data obtained by the CBC measurement performed in the blood cell counting and measuring section 9.
V), etc. are obtained. Further, in the MCU 39, based on the data obtained by the CRP measurement performed in the CRP measurement unit 8, the absorbance change per predetermined time from the calibration curve previously obtained from serum (or plasma) of known concentration, The CRP concentration in whole blood is obtained.

In this case, for CRP measurement, CBC
As in the case of the measurement, since the whole blood to which the anticoagulant is added is used as the sample 3, it is necessary to correct the plasma component volume error caused by using this whole blood. Therefore, in the present invention, the hematocrit value (Hc) is calculated from the RBC (red blood cell count) and the red blood cell volume (MCV) obtained by the CBC measurement.
t) is obtained, and using this hematocrit value, the CRP concentration in whole blood obtained by the CRP measurement is corrected by the following correction equation to obtain the CRP concentration in plasma.

That is, the CRP concentration in whole blood is A
When the hematocrit value is B, the CRP concentration in plasma C
Is calculated by the formula C = A × 100 / (100−B).

The respective measured values obtained by the MCU 39 are stored in a memory built in the MCU 39, for example, and are displayed item by item on the display device 42 or output by the printer 43.

As described above, in the whole blood blood cell immunoassay device of the present invention, CB is generated during the hemolytic and interfering substance removal reactions in the CRP measuring section 8.
Since the blood cell measurement is performed in the C measurement unit 9, the total time for the CRP measurement and the CBC measurement can be shortened, and the result obtained by the CRP measurement described above is corrected by the result obtained by the CBC measurement. Can be done smoothly.

[0040]

According to the whole blood blood cell immunoassay apparatus of the present invention, since the blood cell count and the measurement of the immunity item can be performed simultaneously with the whole blood by one device, the whole blood can be used as the specimen and the serum separation is not necessary. In addition, it is possible to start measurement in a short time after blood collection. Therefore, it is possible to easily and quickly perform the measurement without a specialized inspection engineer.

In the above apparatus, the probe unit section, the arithmetic / control section, the display / output apparatus, etc. can be commonly used for the blood cell count measurement and the immunoassay, and they are individually provided. The cost can be reduced by the amount that can be shared, and the overall structure can be made compact.

[Brief description of drawings]

FIG. 1 is a perspective view showing an example of a whole blood blood cell immunoassay device of the present invention with a side panel removed.

FIG. 2 is a diagram schematically showing an overall configuration of the whole blood blood cell immunoassay device.

FIG. 3 is a flowchart showing an example of a measurement procedure together with FIGS. 4 and 5.

FIG. 4 is a flowchart following the part shown in FIG.

FIG. 5 is a flowchart following the part shown in FIG.

[Explanation of symbols]

8 ... Immunoassay section, 9 ... Blood cell counting and measurement section.

─────────────────────────────────────────────────── ─── Continuation of the front page (58) Fields surveyed (Int.Cl. ⁷ , DB name) G01N 33/53 G01N 33/49

Claims (2)

(57) [Claims] 1. An immunoassay unit for performing an immunoassay and a blood cell count and measurement unit for performing a blood cell count measurement are provided, and the same whole blood sample is used in both of these measurement units, and the result of the immunoassay is obtained by the blood cell count measurement. A whole blood blood cell immunoassay device characterized in that the hematocrit value is used for correction. 2. The whole blood blood cell immunoassay device according to claim 1, wherein blood cell measurement is performed in the blood cell count measurement unit while an immune reaction is caused in the immunoassay unit.